5-HT1A receptor (HTR1A) 5′ region haplotypes significantly affect protein expression in vitro. (18th January 2017)

• Record Type: Journal Article
• Authors: Wu, Xue; Yao, Jun; Ding, Mei; Shi, Zhang-sen; Xu, Feng-ling; Zhang, Jing-jing; Wang, Bao-jie
• Is Part Of: Neuroscience letters. Volume 638(2017)
• Page Start: 51
• Page End: 54
• Publication Date: 2017-01-18
• Subjects: HTR1A -- Polymorphism -- Promoter -- Transcription -- Schizophrenia; Neurology -- Periodicals; Neurology -- Periodicals; Research -- Periodicals; Neurologie -- Périodiques; Neuroanatomie -- Périodiques; Neuropharmacologie -- Périodiques; Neurophysiologie -- Périodiques; Neurology; Periodicals; Electronic journals; 617.48
• DOI: 10.1016/j.neulet.2016.12.011
• Languages: English
• ISSNs: 0304-3940

Abstract

Highlights: A fragment spanning nucleotides −551 to +672 of the HTR1A (Transcription start site +1) 5′ promoter region was used to construct three haplotype plasmids (1-1 to 3-1). The relative fluorescence intensity of plasmids 1-1 and 3-1 was compared to the wild type plasmid 2-1 after transfection of HEK-293 and SK-N-SH cells. In the HEK-293 cells, there was a significant difference between plasmid 1-1(GC) and plasmid 2-1(GG). A significant difference between plasmid 1-1 and plasmid 2-1 was also observed in the SK-N-SH cells. Abstract: The aim of this study was to explore the role of two SNPs (rs6295 & rs113195492) in the HTR1A gene promoter region that regulates the expression of the 5-HT1A receptor. A fragment spanning from −551 to +672 of HTR1A (Transcription start site +1) was cloned into the pGL-3 Basic Vector and three haplotype plasmids composed of two SNPs were constructed. HEK-293 cells and the SK-N-SH cells were transfected with the three plasmids, and the relative fluorescence intensity was measured. In HEK-293 cells, there was a significant difference when the relative fluorescence intensity of plasmid 1-1 was compared to that of plasmid 2-1. However, no significant difference was observed when the luciferase expression of plasmid 2-1 and plasmid 3-1 was analyzed. We also found that the expression trend of the SK-N-SH cells was similar to the HEK-293 cells, but the overall relative fluorescence intensity of the SK-N-SH cells was lower than that of the HEK-293 cells. Our finding showed that the rs6295 SNP, as a suspected variant that indicates susceptibility to schizophrenia, exhibited a higher transcriptional activity. The influence of the rs113195492 locus on schizophrenia needs to be explored further.